Method and compositions producing cherry derived products

ABSTRACT

A method for isolating a mixture of anthocyanins, bioflavonoids and phenolics from cherries using adsorbent resins which are regenerable for reuse is described. The mixture with a consumable carrier is particularly useful in foods and as a dietary supplement or neutraceutical product.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application SerialNo. 60/111,945, filed Dec. 11, 1998, as well as U.S. ProvisionalApplication Serial No. 60/120,178, filed Feb. 16, 1999.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

None

BACKGROUND OF THE INVENTION

(1) Summary of the Invention

The present invention relates to a method of preparation of cherryderived compositions and to a method of use of the compositions derivedfrom the cherries as phytoceutical/neutraceutical dietary supplements oras an additive to foods. In particular, the present invention provides anatural cherry composition containing a mixture of anthocyanins,bioflavonoids and phenolics for use as dietary supplements or as a foodadditive.

(2) Description of Related Art

Many plant-derived compounds may also impart important positivepharmacological or “nutraceutical” traits to foods by way of theirabilities to serve as cellular antioxidants by maintaining low levels ofreactive oxygen intermediates, as anti-inflammatory agents by inhibitingprostaglandin synthesis, or as inhibitors of enzymes involved in cellproliferation. These activities may be important in ameliorating chronicdiseases including cancer, arthritis, and cardiovascular disease(Kinsella et al., Food Tech. 85-89 (1993). Thus, with natural products,the dietary supplement/food industry and nutraceutical companies has theopportunity to employ compounds which can not only enhance foodstability as effectively as synthetic antioxidants, but can also offersignificant health benefits to the consumer.

Cherries are thought to have beneficial health properties in general. Apreferred cherry is Prunus Cerasus L. (Rosacease), cv. MONTMORENCY whichis the major tart cherry commercially grown in the United States. Inorder to challenge the MONTMORENCY monoculture, a new cultivar, BALATONtart cherry (Ujferbertoi furtos), was introduced into the United Statesin 1984, and has been tested in Michigan, Utah, and Wisconsin. BALATONproduces fruits darker than MONTMORENCY.

Colorants like anthocyanins have been regarded as the index of qualityin tart cherries. Most importantly, recent results showed thatanthocyanins such as cyanidin-3-glucoside have strong antioxidantactivities (Tsuda, T., et al, J. Agric. Food Chem. 42:2407-2410 (1994)).The addition of antioxidants is one of the popular methods to increasethe shelf life of food products which is thought to be associated withlipid peroxidation. Natural antioxidants may play an important role inthe prevention of carcinogenesis. Dietary antioxidants may be effectiveagainst the peroxidative damage in living systems (Halliwell, B. and J.M. C. Gutteridge, Free radicals in biology and medicine. OxfordUniversity Press, New York 416-494 (1989); Osawa, T., et al, Role ofdietary antioxidants in protection against oxidative damage. Inantimutagenesis and anticarcinogenesis Mechanisms; Kuroda, Y.; Shankel,D. M., Waters, M. D., Eds.; Plenum Publishing. New York 139-153 (1990)).

Early studies have showed that MONTMORENCY cherry contains theanthocyanins cyanidin-3-gentiobioside and cyanidin-3-rutinoside (Li, K.C., et al., J. Am. Chem. Soc. 78:979-980 (1956)).Cyanidin-3-glucosylrutinoside was also found in six out of the sevensour cherry varieties (Harborne, J. B., et al., Phytochemistry 3:453-463(1964)). Dekazos (Dekazos, E. D., J. Food Sci. 35:237-241 (1970))reported anthocyanin pigments in MONTMORENCY cherry aspeonidin-3-rutinoside, peonidin and cyanidin along withcyanidin-3-sophoroside, cyanidin-3-rutinoside and cyanidin-3-glucoside.However, cyanidin-3-glucosylrutinoside as. well as cyanidin-3-glucoside,cyanidin-3-sophoroside and cyanidin-3-rutinoside were identified as mainpigments in sour cherries. Using HPLC retention values, Chandra et al(Chandra, A., et al., J. Agric. Food Chem. 40:967-969 (1992)) reportedthat cyanidin-3-sophoroside and cyanidin-3-glucoside . were the majorand minor anthocyanins, respectively, in Michigan grown MONTMORENCYcherry. Similarly, cyanidin-3-xylosylrutinoside was detected as a minorpigment in MONTMORENCY cherry (Shrikhande, A. J. and F. J. Francis, J.Food Sci. 38:649-651 (1973)).

In the prior art, production of pure anthocyanins (compounds 1-3 ofFIG. 1) from BALATON and MONTMORENCY cherry juices was carried out firstby adsorbing the pigment on an AMBERLITE XAD-2 (Sigma Chemicals) column(Chandra, A., et al., J. Agric. Food Chem. 41:1062-1065 (1993)). Thecolumn was washed with water until the eluant gave a pH of approximately7.0. The adsorbed pigments along with other phenolics were eluted withMeOH. The resulting crude anthocyanins were fractionated and purified byC-18 MPLC and HPLC, respectively, to afford pure anthocyanins forspectral studies. Purification of 500 mg crude MONTMORENCY anthocyaninsfrom AMBERLITE XAD-2 yielded 60 mg of pure anthocyanins 1-3 compared to391.43 mg from BALATON. This research indicated that crude anthocyaninsfrom MONTMORENCY obtained from the XAD-2 contained a high percentage ofother organic compounds. The AMBERLITE XAD-2 did not allow recycling ofthe resin. There was no attempt to use the crude mixture of phenolicsand anthocyanins for any purpose. U.S. Pat. No. 5,266,685 to Garbutt,U.S. Pat. No. 5,665,783 to Katzakian et al and U.S. Pat. No. 5,817,354to Mozaffar describe various adsorbent resins and their use forunrelated products. These patents are only illustrative of the generalstate of the art in the use of adsorbent resins.

U.S. Patent No. 5,503,867 to Pleva describes the use of whole groundcherries and oat bran in ground meat. The amount of cherries used was 10to 15% by weight and the oat bran is believed to be added to compensatefor the juice in the cherries. In any event, the cherries definitelycontribute a flavor to the meat and the palatability of the product isnot universally accepted.

Recent studies on stabilization of low-fat ground beef with cherrytissue suggest that this plant source contains potent antioxidants whichnot only suppress lipid peroxidation, but also inhibit formation ofheterocyclic aromatic amines and cholesterol oxidation products duringfrying (Gomaa et al., IFT Abstracts No. 68E-7 (1996). The hypothesisused to explain these observations was that polyphenols, such asflavonoids, anthocyanins and anthocyanidins, frequently found in thevacuoles of higher plants such as the cherries were responsible for thisantioxidant effect.

There is a need for natural cherry derived compositions for use,particularly as dietary supplements/nutraceutical or food additives.

SUMMARY OF THE INVENTION

The present invention relates to a method for producing a mixturecomprising anthocyanins, bioflavonoids and phenolics from cherries as acomposition which comprises:

(a) providing an aqueous solution containing the anthocyanins,bioflavonoids and phenolics from the cherries;

(b) removing the anthocyanins, bioflavonoids and phenolics onto a resinsurface from the aqueous solution;

(c) eluting the resin surface with a eluant to remove the anthocyanins,bioflavonoids and phenolics from the resin surface; and

(d) separating the eluant from the anthocyanins, bioflavonoids andphenolics.

Further, the present invention relates to a method for producinganthocyanins, bioflavonoids and phenolics from cherries as a compositionwhich comprises:

(a) providing a first batch of cherries, wherein the cherries are freshor quick frozen and thawed;

(b) disrupting the cherries and separating pulp from the juice;

(c) extracting the anthocyanins, bioflavonoids and phenolics from thepulp into an aqueous solution;

(d) removing the anthocyanins, bioflavonoids and phenolics ontoadsorbent resin particles from the aqueous solution containing theanthocyanins, bioflavonoids and phenolics separated from the pulp;

(e) washing the resin particles with a lower alkanol to remove theanthocyanins, bioflavonoids and phenolics from the resin particles;

(f) separating the alkanol from the anthocyanins, the bioflavonoids andphenolics; and

(g) repeating steps (a) to (e) with the separated alkanol and the resinparticles from which the anthocyanins, bioflavonoids and phenolics havebeen removed with a second batch of the cherries.

Further, the present invention relates to a consumable composition whichcomprises in admixture:

(a) dried mixture of isolated anthocyanins, bioflavonoids and phenolicsfrom cherries; and

(b) a food grade carrier, wherein the weight ratio of (a) to (b) isbetween about 0.1 to 100 and 100 to 0.1.

Finally, the present invention relates to a method for feeding a mammalwhich comprises:

feeding the mammal a consumable composition which comprises inadmixture:

(a) dried mixture of isolated anthocyanins, bioflavonoids and phenolicsremoved from cherries; and

(b) a food grade carrier wherein the weight ratio of (a) to (b) isbetween about 0.1 to 100 and 100 to 0.1. It, is preferred that thecomposition contain at least in part dried cherry pulp.

The term “anthocyanins” means the compounds that impart color incherries.

The term “bioflavonoids” means the isoflavonoids and flavonoid compoundscontained in cherries.

The term “phenolics” refers to compounds with a phenyl group and havingone or more hydroxyl groups from cherries.

OBJECTS

It is therefore an object of the present invention to provide a naturalsource cherry composition which can be used in foods or as dietarysupplements or nutraceuticals. Further, it is an object of the presentinvention to provide a method for isolating the composition on acommercial scale. Finally, it is an object of the present invention toprovide a natural source composition which is economical to prepare andeasy to use. These and other objects will become increasingly apparentby reference to the following description and the drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the structure of the isolated anthocyanins (colorants) fromBALATON and MONTMORENCY cherries. The aglycone cyanidin has a hydroxylgroup at position 3.

FIGS. 2 and 3 are drawings showing the major bioflavonoids isolated fromthe cherries, as described in provisional application Serial No.60/111,945, filed Dec. 11, 1998.

FIG. 4 shows the phenolics isolated from tart cherries.

FIG. 5 shows the steps in the method of the present invention asdescribed in Examples 1 and 2.

FIG. 6 is a schematic drawing showing the use of an open vessel 10 forholding resin beads, which remove anthocyanins and phenolics from thecherry juice.

DESCRIPTION OF PREFERRED EMBODIMENTS

The cherries used in the present invention can be sweet or sour,although the latter are preferred since they contain high levels ofmalic acid in addition to other organic acids which contributes to thesour taste of tart cherries. The method of the present inventionisolates malic acid and other organic acids containing sugars which canbe used in foods to provide tartness and flavor. Most preferred are theBALATON and MONTMORENCY cherries.

The isolated mixture of anthocyanins, bioflavonoids and phenolics can betableted and used as a natural nutraceutical/dietary supplement. Ingeneral, the tablets provide a daily dose of the anthocyanins andbioflavonoids of about 1 to 200 mg, preferably a daily dose of 60-100mg. One hundred (100) cherries provide 60 to 100 mg of anthocyanins. Thephenolics (FIG. 4) are provided in an amount of 0.1 to 50 mg as a dailydose. One hundred cherries provide 1-50 mg of phenolics. The amount ofthe anthocyanins, bioflavonoids and phenolics can be adjusted byisolating the individual compounds and blending them-together. It ispreferred to use the natural mixture of the anthocyanins, bioflavonoidsand phenolics which is isolated by the resin.

The resin has a surface to which the anthocyanins, bioflavonoids and thephenolics are adsorbed. A preferred class of adsorptive resins arepolymeric crosslinked resins composed of styrene and divinylbenzene suchas, for example, the AMBERLITE series of resins, e.g., AMBERLITE XAD-4and AMBERLITE XAD-16, which are available commercially from Rohm & HaasCo., Philadelphia, Pa. Other polymeric crosslinked styrene anddivinylbenzene adsorptive resins suitable for use according to theinvention are XFS-4257, XFS-4022, XUS-40323 and XUS-40322 manufacturedby The Dow Chemical Company, Midland, Mich., and the like.

It is preferred to use commercially available, FDA-approved,styrene-divinyl-benzene (SDVB) cross-linked copolymer resin, (e.g.,AMBERLITE XAD-16). Thus, in the preferred embodiment, AMBERLITE XAD-16,commercially available from Rohm and Haas Company, and described in U.S.Pat. No. 4,297,220, herein incorporated by reference, is used as theresin. This resin is a non-ionic hydrophobic, cross-linked polystyrenedivinyl benzene adsorbent resin. AMBERLITE XAD-16 has a macroreticularstructure, with both a continuous polymer phase and a continuous porephase. In a particularly preferred embodiment, the resin used in thepresent invention has a particle size ranging from 100-200 microns.

It is contemplated that other adsorbents such as those in the AMBERLITEXAD adsorbent series which contain hydrophobic macroreticular resinbeads, with particle sizes in the range of 100-200 microns, will also beeffective in the methods of the present invention. Moreover, differentvariations of the AMBERLITES, such as the AMERCHROM CG series ofadsorbents, used with particle sizes in the range of 100-200 microns,may also be suitable for use in the present invention. The AMBERLITEXAD-16 is preferred since it can be re-used many times (over 100 times)However, it is contemplated that for food, the use ofgovernmentally-approved resins in the present invention will beconsidered important and/or desirable.

Any solvent can be used to remove the adsorbed anthocyanins,bioflavonoids and phenolics. Preferred are lower alkanols containing 1to 4 carbon atoms and most preferred is ethanol (ethyl alcohol) since itis approved for food use. Typically the ethanol is azeotroped withwater; however, absolute ethanol can be used. Water containing malicacid and sugars in the cherries pass through the column. These arecollected and can be used in foods as flavors.

The anthocyanins, bioflavonoids and phenolics are preferably isolatedfrom the BALATON and the MONTMORENCY cherries. The composition of thecherries is in part shown in part by U.S. application Ser. No.08/799,788 filed Feb. 12, 1997 and in part U.S. application Ser. No.60/111,945, filed Dec. 11, 1998 which are incorporated by referenceherein.

The term “carrier” or “bulking agent” is used to mean a compositionwhich is added to increase the volume of the composition of the purifiedcomposition from the cherry. Preferred is dried cherry pulp. Theseinclude any edible starch containing material, protein, such as non-fatdry milk. Within this group are flour, sugar, soybean meal, maltodextrinand various condiments, such as salt, pepper, spices and herbs, forinstance. The bulking agent is used in an amount between about 10⁻⁶ and10⁶ parts by weight of the mixture.

The composition is introduced into-the food in an amount between about0.1 and 10 mg/gm of the active ingredients of the food. The amount ispreferably selected so as to not affect the taste of the food and toproduce the most beneficial result. The food can be high (wet) or lowmoisture (dry) as is well known to those skilled in the art. When usedas a dietary supplement the tablets contain between 0.1 to 1 gram ofactive ingredient.

Methods have been developed for extraction and isolation ofphytochemicals (Chandra, A., et al., J. Agric. Food Chem. 41:1062(1992); Wang, H., et al., J. Agric. Food Chem. 45:2556-2560 (1997)) andfor rapid screening of antioxidant activity (Arora, A. and G. M.Strasburg, J. Amer. Oil Chem. Soc. 74:1031-1040 (1997)). These methodsare being utilized to identify and characterize the antioxidantcompounds from BALATON and MONTMORENCY cherries. Juiced cherry tissuewas sequentially extracted with hexane, ethyl acetate and methanol. Bothmethanol and ethyl acetate fractions showed strong antioxidant activityin the screening assay. The ethyl acetate fraction was further purifiedby silica gel vacuum liquid chromatography to yield four subfractions;the subfraction which showed the strongest antioxidant activity wasfurther separated into seven fractions by preparative reverse phaseHPLC. FIGS. 2 and 3 show the bioflavonoids isolated from the BALATONcherries. There are thus numerous analogous or homologous compounds inthe tart cherries.

Two novel phenolic compounds were identified: I) 1-(3′-4′-dihydroxycinnamoyl)-2,3-dihydroxy cyclopentane, and II) 1-(3′-4′-dihydroxycinnamoyl)-2,5-dihydroxy cyclopentane. Other compounds isolated from theethyl acetate extract of cherry fruits and characterized by spectralmethods include: 1-(3′-methoxy, 4′-hydroxy cinnamoyl) quinic acid,2-hydroxy-3-(2′-hydroxyphenyl) propanoic acid, methyl2-hydroxy-3-(2′-hydroxyphenyl) propanoate, D(+)-malic acid, β-sitosterolad β-sitosterol glucoside. FIG. 4 shows some of the phenolics which wereisolated. The anthocyanin components obtained from the juice fractionalso have been identified and fully characterized (Chandra, A., et al.,J. Agric. Food Chem. 41:1062 (1992); Wang, H., et al., J. Agric. FoodChem. 45:2556-2560 (1997); the results indicate that these compoundscontain potent antioxidant activity.

EXAMPLES 1 and 2

As shown in FIG. 5, individual quick frozen (IQF) cherries (which hadbeen pitted) were defrosted and blended in an industrial WARING blender.The mixture was centrifuged at 10,000 rpm and the juice was decanted.The residue, pulp, was further pressed with cheese cloth to remove anyadditional juice.

The pulp was lyophilized at 15° C. The juice was processed on AMBERLITEXAD-16 HP resin to produce cherry sour, anthocyanins, bioflavonoids andphenolics. The XAD-16 resin, 1 kg, was washed with ethanol (1-2 L) andthen washed with water (6 L). The XAD-16 resin was allowed to stand inwater for 1 hour before loading into a glass column (10 ID×90 cm long)with a cotton plug. The packed column was washed with water (2 L) beforeloading the juice for separation. 800 mL juice was purified each time.The juice was added onto the surface of the column and allowed to settlewith no flow. It was then eluted with water and the first 1 L wasdiscarded. The next 2 L of washing was collected, since it contained thecherry juice which was sour since it contained malic acid and sugarsfrom the cherries. The column was then washed with an additional 4 L ofwater in the case of BALATON and 5 L for MONTMORENCY cherry juice. Oncethe cherry juice was collected, the remainder of the washing with waterwere discarded. The column was then eluted with ethanol (1.3-1.5 L) andcollected the red solution containing anthocyanins, bioflavonoids andphenolics (700-800 ml). The column was then run dry and washed with 10 Lof water before repeating the process many of times (over 100).

The red alcoholic solution was then evaporated under vacuum a (20millitorr) to remove ethanol and the aqueous solution, stabilized with50 ppm ascorbic acid, was lyophilized at 10° C. The red powder wascollected and stored.

Example 1 results: BALATON cherry Weight of IQF cherries 15.74 kg Weightof dried pulp 605 g Volume of juice 12.16 L Weight of anthocyanins,bioflavonoids 31.35 g and phenolics (red powder) Volume of sourbyproduct @ 35 L (malic acid and sugars) Example 2 results: MONTMORENCYcherry Weight of IQF cherries 30.45 kg Weight of dried pulp 895 g Volumeof juice 24.03 L Weight of anthocyanins, bioflavonoids 47 g andphenolics (red powder) Volume of cherry by-product @ 75 L (malic acidand sugars)

The red powders of Examples 1 and 2 were preferably mixed with driedpulp as a carrier and tableted into 1 to 1000 mg tablets including thecarrier (1 adult daily dose).

Various food grade acids can be added to the isolated anthocyanins,bioflavonoids and phenolics to prevent decomposition. Preferably they donot add flavor. Ascorbic acid (vitamin C) is preferred. The acid can beadded before or after the drying of the cherry compounds.

For small scale processing, lyophilization is used to remove water. Forlarger scale production, drying in an air circulating oven is preferred.

EXAMPLE 3

As shown in FIG. 6, an open vessel 10 is provided with an inlet line 11and an, outlet line 12, with valves 13 and 14, respectively. The resinbeads 15 are provided in the open vessel 10. Water is introduced intothe vessel 10 and then removed through outlet line 12 and discarded. Thecherry juice (without the pulp or pits) as in Example 1 is introduced tothe vessel 10 and allowed to stand for 25 minutes. The temperature ofthe water and juice is between about 20 and 30° C. The cherry juiceresidue containing malic acid and sugars is then removed through theoutlet line 12 and retained as a food flavoring. The resin 15 in thevessel is then washed again with water from inlet line 11 and thenremoved and discarded through outlet line 12. The anthocyanins,bioflavonoids and phenolics on the resin particles are then extractedusing 95% ethanol introduced through inlet line 11. The ethanolcontaining the anthocyanins, bioflavonoids and phenolics is removed fromthe vessel 10. The ethanol is removed from the anthocyanins,bioflavonoids and phenolics and dried using flash drying under nitrogen.The resulting powder is preferably then mixed with dried cherry pulp orother carrier as in Example 1. The resin particles are washed with waterand then the resins and ethanol are recycled many times.

EXAMPLE 4

Crude ethyl acetate extracts from cherries (containing anthocyanins,bioflavonoids and phenolics) were tested in aqueous solution undervarious conditions using a fluorescent assay for antioxidant activity.The fluorescent assay is described first.

Fluorescence Assay for Antioxidant Activity (general): The need toscreen large numbers of compounds or extracts for antioxidant activityrequires that a model system (or systems) be employed which reasonablywell represents the structural and functional characteristics of thecomposition alone or in the food product. The test must also besensitive, rapid, and inexpensive. A fluorescence-based assay forevaluating antioxidant efficacy was used (Arora, A., and G. M.Strasburg, J. Am. Chem. Soc. 1996)). Large unilamellar vesiclesconsisting of 1-stearoyl-2-linoleoly-sn-glycero-3-phosphocholine wereprepared, which closely resemble the properties of biological membranes,one of the primary sites of peroxidation. A fluorescent probe,1,6-diphenylhexatriene propionic acid, is incorporated into themembranes such that the polar head group anchors the probe near theaqueous interface, while the hydrophobic portion lies parallel to thefatty acid chains. This probe reacts with the free radicals generatedduring peroxidation, resulting in a decrease in fluorescence intensitywith time. A peroxidation initiator (such as ferrous metal ions or thefree radical generator AAPH (Azobis-[2-amidino propane hydrochloride])is used to start the reaction, and the kinetics of fluorescence decreaseare determined in the presence or absence of the antioxidant compositionto be tested. An assay for a compound at a given concentration presentlytakes only twenty-one minutes, consumes only a few micrograms of lipid,and can be readily conducted with a simple fluorometer.

Large unilamellar vesicles (LUVs) were prepared from1-stearoyl-2-linoleoyl-sn-glycero-3-phosphocholine according to theprocedure outlined by MacDonald et al (MacDonald, R. C., et al.,Biochim. Biophys. Acta 1061:297-303 (1991)). Briefly, the lipid wasdissolved in chloroform, and was dried to a thin film using a rotaryevaporator. The dried film was resuspended in an aqueous buffer, and wasrepeatedly extruded through a polycarbonate filter of 100 nm pore sizeusing a Liposofast piposome extruder (Avestin; Inc., Ottawa, Canada).The homogeneity of size (80-100 nm) and the unilamellar nature of thevesicles were confirmed using freeze-fracture scanning electronmicroscopy. The fluorescent probe, diphenylhexatriene-propionic acid(DPH-PA), was incorporated into the vesicles during preparation at amole ratio of 1:350 (probe:lipid). For the fluorescence experiments,LUVs containing DPH-PA is suspended at a final concentration of 100 μMin 100 mM NaCl, 50 mM tris-HEPES buffer at pH 7.0. The fluorescent probewas excited at 384 nm and emission was monitored at 423 nm. Lipidoxidation is inhibited in the LUVs by addition of ferrous ions or thefree radical generator AAPH; the progress was monitored by the decreaseof the fluorescence intensity of DPH-PA resulting from reaction withfree radicals generated over twenty-one minutes. A plot of the decreaseof fluorescence intensity as a function of time was used to determinethe kinetics of lipid oxidation. The results show that a mixture of thecrude anthocyanin extract with ethylacetate was effective in inhibitingoxidation.

Solvent extraction of the anthocyanins, bioflavonoids and phenolics canbe used; however this is not preferred where the product is to be usedas a food and for expense reasons. Where the preferred adsorbent resinsare used, this step is unnecessary. It is also possible to separate andrecombine the components using chromatography; however, for the purposeof the present invention, this is far too expensive since it involveshigh pressure liquid chromatography.

EXAMPLE 5

The compositions were tested for anti-inflammatory activity usingcyclooxygenase I and II (COX-I and COX-II) in an assay as described inWang et al., J. Nat. Products 62:294-296 (1999); Wang et al., J. of Ag.and Food Chemistry, 47: 840-844 (1999) and Wang et al., J. of Nat.Products, 62:86-88 (1999). The results were that the compositionsexhibited anti-inflammatory activities, specifically strong inhibitionof COX-I and COX-I.

It is intended that the foregoing description be only illustrative ofthe present invention and that the present invention be limited only bythe hereinafter appended claims.

I claim:
 1. A method for producing a mixture comprising anthocyanins,bioflavonoids and phenolics from cherries as a composition whichcomprises: (a) providing an aqueous solution containing theanthocyanins, bioflavonoids and phenolics from the cherries; (b)removing each of the anthocyanins, bioflavonoids and phenolics onto aresin surface from the aqueous solution, wherein the resin is for usewith food products; (c) eluting the resin surface with an eluant toremove the anthocyanins, bioflavonoids and phenolics as the mixture fromthe resin surface; and (d) separating the eluant from the anthocyanins,bioflavbnoids-and phenolics to produce the mixture, wherein the methodis effective to produce a mixture containing two phenolics of theformula:

where R₁ and R₂ are different and selected from the group consisting ofH and OH; (e) repeating steps (a) to (d) with the separated eluant andthe resin particles from which the anthocyanins, bioflavonoids andphenolics have been removed with multiple batches of the cherries. 2.The method of claim 1 wherein the anthocyanins, bioflavonoids andphenolics are mixed with a pulp from the cherries and then are dried. 3.The method of claim 2 wherein in addition the mixture of theanthocyanins, the bioflavonoids, the phenolics and the pulp are formedinto a tablet.
 4. The method of claim 1 wherein the anthocyanins,bioflavonoids and phenolics are dried and then mixed with a dried cherrypulp.
 5. The method of claim 1 wherein the resin is a polymericcross-linked styrene and divinylbenzene adsorptive resin.
 6. The methodof claim 5 wherein the resin surface is a macroreticular structure witha continuous polymer phase-and a continuous pore phase.
 7. The method ofclaim 6 wherein the resin surface is as particles having a size betweenabout 100 to 200 microns.
 8. A method for producing a mixture ofanthocyanins, bioflavonoids and phenolics from cherries as a compositionwhich comprises: (a) providing a first batch of cherries, wherein thecherries are fresh or quick frozen and thawed; (b) disrupting thecherries and separating pulp from the juice; (c) extracting theanthocyanins, bioflavonoids and phenolics from the pulp into an aqueoussolution; (d) removing each of the anthocyanins, bioflavonoids andphenolics as a mixture onto adsorbent resin particles from the aqueoussolution containing the anthocyanins, bioflavonoids and phenolicsseparated from the pulp, wherein the resin particles are for use withfood products; (e) washing the resin particles with a lower alkanol toremove the anthocyarnins, bioflavonoids and phenolics as the mixturefrom the resin particles; (f) separating the alkanol from theanthocyanins, the bioflavonoids and phenolics as the mixture, whereinthe method is effective to produce a mixture containing two phenolics ofthe formula:

where R₁ and R₂ are different and selected from the group consisting ofH and OH; and (g) repeating steps (a) to (e) with the separated alkanoland the resin particles from which the anthocyanins, bioflavonoids andphenolics have been removed with multiple batches of the cherries. 9.The method of claim 8 wherein the alkanol is ethanol.
 10. The method ofany one of claims 8 or 9 wherein the cherries are individually quickfrozen.
 11. The method of any one of claims 8 or 9 wherein the resinparticles are in the form of a column.
 12. The method of any one ofclaims 8 or 9 wherein the cherries are tart cherries.
 13. The method ofany one of claims 8 or 9 wherein the anthocyanins, bioflavonoids andphenolics are mixed with the pulp from the cherries and then are dried.14. The method of any one of claims 8 or 9 wherein the anthocyanins,bioflavonoids and phenolics are dried and then mixed with a dried cherrypulp.
 15. The method of any one of claims 8 or 9 wherein in addition theanthocyanins, bioflavonoids and phenolics are mixed with a cherry pulpand then are dried and wherein the mixture of the anthocyanins, thebioflavonoids, the phenolics and the pulp are then formed into a tablet.16. The method of any one of claims 8 or 9 wherein the anthocyanins,bioflavonoids and phenolics are dried and then mixed with dried pulp andwherein the mixture of the anthocyanins, the bioflavonoids, thephenolics and the pulp are then formed into a tablet.
 17. A method forproducing a mixture comprising anthocyanins, bioflavonoids and phenolicsfrom cherries with a bulking agent for use as a nutraceuticalcomposition which comprises: (a) providing an aqueous solutioncontaining the anthocyanins, bioflavonoids and phenclics from thecherries; (b) removing each of the anthocyanins, bioflavonoids andphenolics as a mixture onto a resin surface from the aqueous solution;(c) eluting the resin surface with an eluant to remove the anthocyanins,bioflavonoids and phenolics as the mixture from the resin surface; (d)separating the eluant from the anthocyanins, bioflavonoids and phenolicsto provide the mixture, whereby the method is effective to produce amixture containing two phenolics of the formula:

where R₁ and R₂ are different and selected from the group consisting ofH and OH; and (e) combining the mixture with a bulking agent and forminga dosage unit so that the mixture with the bulking agent provides anamount of the mixture which produces the nutraceutical composition. 18.The method of claim 17 wherein the mixture of anthocyanins,bioflavonoids and phenolics are-mixed with a pulp from the cherries asthe bulking agent in step (g) and then are dried.
 19. The method ofclaim 17 wherein the anthocyanins, bioflavonoids and phenolics are driedafter step (f) and then mixed with a dried cherry pulp as the bulkingagent.
 20. The method of claim 17 wherein in addition the mixture of theanthocyanins, the bioflavonoids, the phenolics and the pulp as thebulking agent are formed into a tablet as the dosage unit.
 21. A methodfor producing a mixture of anthocyanins, bioflavonoids and phenolicsfrom cherries with a bulking agent for use as a nutraceuticalcomposition which comprises: (a) providing a first batch of cherries,wherein the cherries are fresh or quick frozen and thawed; (b)disrupting the cherries and separating pulp from the juice; (c)extracting the anthocyanins, bioflavonoids and phenolics from the pulpinto an aqueous solution, wherein the resin is for use with foodproducts; (d) removing each of the anthocyanins, bioflavonoids andphenolics as a mixture onto adsorbent resin particles from the aqueoussolution containing the anthocyanins, bioflavonoids and phenolicsseparated from the pulp; (e) washing the resin particles with a loweralkanol to remove the anthocyanins, bioflavonoids and phenolics as themixture from the resin particles; (f) separating the alkanol from theanthocyanins, the bioflavonoids and phenolics to provide the mixture,whereby the method is effective to produce a mixture containing twophenolics of the formula:

where R₁ and R₂ are different and selected from the group consisting ofH and OH; and (g) comnbining the mixture with a bulking agent andforming a dosage unit so that the mixture with the bulking agentprovides an amount of the mixture which produces the nutraceuticalcomposition.
 22. The method of claim 21 wherein the alkanol is ethanol.23. The method of any one of claims 21 or 22 wherein the cherries areindividually quick frozen.
 24. The method of any one of claims 21 or 22wherein the resin particles are in the form of a column.
 25. The methodof any one of claims 21 or 22 wherein the cherries are tart cherries.26. The method of claim 21 wherein the anthocyanins, bioflavonoids andphenolics are mixed with the pulp from the cherries and then are dried.27. The method of claim 21 wherein the anthocyanins, bioflavonoids andphenolics are dried and then mixed with a dried cherry pulp.
 28. Themethod of claim 21 wherein in addition the mixture of the anthocyanins,the bioflavonoids, the phenolics and the pulp as the bulking agent areformed into a tablet as the dosage unit.